Application of competitive indirect enzyme-linked immunosorbent assay (Ci-ELISA) for monitoring the degree of frozen denaturation of bovine myosin

The denaturation of myosin on freezing and frozen storage was monitored using competitive indirect enzyme-linked immunosorbent assay (Ci-ELISA) formatted with polyclonal antibodies anti-MWM IgG, anti-S-1 IgG and anti-LMM IgG raised against the antigens (Ags) bovine myosin whole molecules (MWMs), heavy meromyosin S-1 (myosin head part, S-1) and light meromyosin (myosin tail part, LMM) respectively. Beef slices and cuts stored at -20 degrees C or -50 degrees C lost immune affinity with all antibodies, in particular anti-LMM IgG. Repeated thawing-refreezing treatment caused more myosin denaturation than simple freezing. Myosin from beef stored at -20 degrees C was denatured more than that stored at -50 degrees C. The immune affinities between anti-LMM IgG and thawed samples were similar to those from anti-MWM IgG. We were unable to differentiate reliably between fresh and thawed beef using anti-S-1 IgG. Myosin was denatured by freezing, in particular its tail part (LMM).